Solid-phase method of production of water-soluble bioactive nanocomposite based on hyaluronic acid modified by citric acid and gold nanoparticles

FIELD: biotechnology.

SUBSTANCE: method of production of the water-soluble bioactive nanocomposite comprising salt of hyaluronic acid modified by citric acid or salt of citric acid, as a matrix, and gold nanoparticles as the filler is carried out by chemical interaction of solid-phase powders of salt of hyaluronic acid, citric acid or salt of citric acid and aurichlorohydric acid or salt of gold under the temperature from -18° to 125°C, under conditions of simultaneous action of pressure from 50 to 1000 MPa and the shear deformation in a mechanochemical reactor.

EFFECT: invention enables to obtain water-soluble bioactive nanocomposite with reliably predictable characteristics, namely the high yield of the target product, high gold content, controlled size of gold nanoparticles, narrow distribution by size of nanoparticles of gold, significant increase in resistance of the composite during long storage the above parameters of the composite are maintained for at least one year, the method is carried out in the absence of the liquid medium and does not require the stage of purification and concentration.

20 cl, 18 ex

 

The invention relates to natural polymers from the class of polysaccharides, namely solid-phase method for producing a water-soluble bioactive nanocomposites nanocomposite based on modified citric acid (sa) or a salt of citric acid salt of hyaluronic acid (ha) and gold nanoparticles (NPS gold), which may find application in medicine, in particular the photon capture therapy (ERT), photothermal therapy, photo - and radiosensibility, chemotherapy, treatment of rheumatoid arthritis, antovic therapy, cosmetology, aesthetic dermatology and plastic surgery.

A method of obtaining nanoparticles of noble metals and the manufacture of materials and devices containing nanoparticles (patent RU 2233791, 2004.08.10). This method of producing nanoparticles includes forming a two-phase system - molecular layer on the surface of the aqueous phase containing the water-insoluble ORGANOMETALLIC precursor molecules (used connection - acetate, palladium, Au(P(C6H5)3)Cl), and processes for the synthesis of metal nanoparticles by chemical transformations of the source reagent precursor under the action of chemical effects or chemical and physical effects, or combinations thereof in a monomolecular layer on the surface of the liquid phase. When the reducing agent (borohydride sodium) was administered during the pilot phase. The method of manufacture of materials containing nanoparticles, is the introduction of the above particles in the material.

The invention is known from RU 2383554, 10.03.2010 is the connection on the basis of the Ledger, where the alcohol group GK tarifitsirovana Reina in free form or in the form of a derived pharmaceutically active compounds in vivo, and where the acid group Reina able to form the ester linkage with a hydroxyl group of hyaluronic acid or its pharmaceutically acceptable salt.

You know the solution CN 102552309, 11.07.2012. The invention relates to the field of synthesis of new medicines for the treatment of tumors, in particular to a method to obtain zolotodolarovoho acid and its application. The invention consists in obtaining a solution of hyaluronic acid or sodium hyaluronate and then adding the halide of gold and obtaining a homogeneous solution. The molar ratio of GC:gold = 1:1. In the resulting solution is injected ethanol, cooled at 0°C, with the aim of vicadine the target product. The precipitate (target product) is filtered off and dried under vacuum at -40°C. of the Target product is stored in a dry form. The invention provides the use of gold hyaluronic acid in the preparation of drugs for the treatment of tumors, in particular provides the use of Salathiel ronojoy acid in the preparation of drugs for the treatment of cancer, and more specifically involves the application of zolotodolarovoho acid in the preparation of the treatment of liver cancer. Zolotogolovuyu acid has the effect of inhibition of various cancer cells, in particular, has a significant effect on the suppression of liver cancer cells.

Known solid phase method for the production of bioactive nanocomposites nanocomposite based on modified sulfur-containing compounds crosslinked hyaluronic acid salt and gold nanoparticles [see patent RU 2416389, 20.04.2011]. The disadvantages of the method include: the preliminary stage of obtaining crosslinked hyaluronic acid salt in the form of a film using a number of cross-linking agents from the class of esters, the method combines the receipt of the modified GC in a solid coated WOOFER noble metal in the gas phase. In this way, in contrast to the claimed invention, the synthesis of NPS gold is due to the action of electric current of high voltage; the introduction of sulfur-containing compounds aimed at the modification of ha in the presence of crosslinking agents.

Thus, the prior art is not known solid phase method of obtaining water-soluble bioactive nanocomposites nanocomposite based on salts of hyaluronic acid, a chemically modified with citric acid or its salt, and gold nanoparticles.

Object of the present invention is to provide an environmentally safe is wow, a fundamentally new method of obtaining water-soluble bioactive nanocomposites nanocomposite comprising a modified citric acid or its salt salt of hyaluronic acid as a matrix and the gold nanoparticles as a filler, in one technological mode in the absence of a liquid medium, without a lot of energy, labor and vadastra, and obtaining the target product with a high yield and a high gold content.

The problem is solved in that created a fundamentally new environmentally friendly method of obtaining water-soluble bioactive nanocomposites nanocomposite comprising a modified citric acid or a salt of citric acid salt of hyaluronic acid as a matrix and the gold nanoparticles as a filler. How is that carried out the chemical interaction of solid powders, salts of hyaluronic acid, citric acid or salts of citric acid and soloconsolidation acid or gold salts at temperatures from -18° to 125°C, under simultaneous pressure from 50 to 1000 MPa and shear strain in mechanochemical reactor. The degree of filling of the composite gold (with oxidation States 0, +2, +3) is from 5·10-6-5·10-1wt. % to 7 wt. %. The nanoparticles have a size of from 1 to 20 nm.

As the salt of hyaluronic acid COI is lsout salt, selected from the range: tetraalkylammonium, lithium, sodium, potassium, calcium, magnesium, barium, zinc, aluminum, copper, gold or mixed salt of hyaluronic acid of the above number or Hydrosol hyaluronic acid.

In particular, the salt of hyaluronic acid is a sodium salt or a mixed salt or hydrometridae salt.

The modifier is citric acid (2-hydroxy-1,2,3-propanetricarboxylate acid, 3-hydroxy-3-carboxypentyl acid) with the gross formula C6H8O7or salts of citric acid - citrate (sodium, potassium, magnesium and so on).

In the stabilizer is used as glutathione or cysteine, which is optionally injected into the reactor. Stabilizers (in particular, glutathione or cysteine) - chemicals that carry SH - group and organized in a monolayer on the particle surface through the formation of covalent bonds between sulfur atoms and gold.

Glutathione is a Tripeptide γ-glutamylcysteinylglycine (2-amino-5-{[2-[(carboxymethyl)amino]-1-(mercaptomethyl)-2-oxoethyl]amino}-5-oxopentanoic acid).

As cysteine using L - and D - isomers of α-amino-β-mercaptopropionate (thiopropionate) acid corresponding to the formula HO2CCH(NH2)CH2SH.

As gold reagents used soloconsolidation Ki the lot (HAUCl 4·nH2O, where n=3 or 4), and gold salts - aurate: solutocapillary potassium (K[AuI4]·nH2O, n=3 or 4), tetrachloroaurate(III) potassium (K[AuCl4] contains 52% of gold), tetrachloroaurate(III) ammonium (NH4[AuCl4] contains 52% of gold), tetrachloroaurate(III) sodium (Na[AuCl4]·n H2O, contains 49% of gold), tetrabromophenol(III) sodium (Na[AuBr4] contains 33% gold), dicyanoaurate potassium (K[Au(CN)2] contains 68.2% of gold), tetracenomycin(III) potassium (K[Au(CN)4], contains 58% gold), disulfiram(I) ammonium(NH4)3[Au(SO3)]2contains 10% gold), bis(thiosulfate)Aurat(I) sodium (Na3[Au(S2O3)2]·H2O, contains 37%), chloro(triphenylphosphine)gold(I) ([AuCl(PPh3)], contains 39% of gold).

The molar ratio of the salt of hyaluronic acid to citric acid is from 1:0.01 to 1:20, respectively.

The molar ratio of soloconsolidation acid to citric acid is in the range from 1:1 to 1:20, respectively.

The molar ratio of zolotoizvlekatelnoj potassium to citric acid is in the range from 1:1 to 1:20, respectively.

The molar ratio of soloconsolidation acid for glutathione is in the range from 1:1 to 1:15, respectively.

The molar ratio of soloconsolidation acid to cysteine is in the range from 1:1 to 1:20 with the responsibility.

The molar ratio of zolotoizvlekatelnoj potassium to the glutathione is in the range from 1:1 to 1:15, respectively.

The molar ratio of zolotoizvlekatelnoj potassium to the cysteine is in the range from 1:1 to 1:20, respectively.

The duration of the effects of pressure and shear strain, in particular, is in the range from 0.1 to 60 minutes, in particular 6 minutes at a pressure of 1000 MPa. As a mechanochemical reactor can be used, in particular, Bridgman anvils or machine screw type.

In the case of the implementation process, where mechanochemical reactor are Bridgman anvils, the reaction mixture is subjected to shear by changing the angle of rotation of the lower anvil, in particular in the range from 50 to 350 degrees. For better implementation of the method, preferably the original reagents pre-homogenized in the mixer at a temperature from -18 to 5°C until a homogeneous powder mixture. In this case you can use as a mixer mill or a mixer, a screw type, for example twin-screw extruder, a fluidized bed apparatus.

In particular, mechanochemical reactor device is a screw type, for example, selected from the set: twin screw extruder with a unidirectional rotation of the screw, twin-screw extruder with protivopolojna the directional rotation of the augers, twin screw extruder with a set of Cams of different types, such as transport, locking, peretiraya.

Preferably, if the method will be implemented sequentially, for example, first carry out a chemical reaction of the salt of hyaluronic acid with citric acid or its salt, after which the product is subjected to interaction with ZHUK, then if necessary add the calculated amount of glutathione or cysteine.

In the reaction mixture optionally, you can enter at least one stabilizing additive. The stabilizing additive is a high molecular weight polymer additive that reduces the degree of aggregation of previously chemically stable bass gold when translating nanocomposites nanocomposite in solution due to passive adsorption of the polymer on the particle surface. The stabilizing additive is introduced into a mechanochemical reactor at the stage of obtaining bioactive nanocomposites nanocomposite solid (solid phase).

As stabilizing additives used substance selected from the group of: carboxymethylcellulose (CMC), tetraalkylammonium, lithium, sodium, potassium, calcium, magnesium, barium, zinc, aluminum, copper, gold or mixed salt of CMC from the above number or Hydrosol CMC, hydroxyethyl cellulose, hydroxypropylcellulose. Moreover, the molar ratio of the group of: salt of hyaluronic acid for stabilizing additive in the range from 100:1 to 1:1.

The invention allows to obtain a water-soluble bioactive nanocomposite with reliably predictable characteristics, namely high yield of the target product (to 99.9%); high gold content (up to 7 wt. %); controlled by the amount of bass from 1 to 20 nm; a narrow distribution in size LF in gold content in the composition 5-7 wt. % the percentage WOOFER of the same size up to 70% and if the gold content <5 wt. % to 92%. Achieved a significant increase in resistance of the composite during prolonged storage. The above parameters of the composite are stored for at least a year. Method for the production of bioactive nanocomposites nanocomposite is carried out in the absence of a liquid medium and does not require a stage of purification and concentration.

When implementing the proposed method to obtain water-soluble bioactive nanocomposites nanocomposite occur following processes:

1) at temperatures from -18°C to 5°C, a pressure of from 50 to 100 MPa and duration of the reaction up to 15 minutes, grinding and amorphization biopolymer substrate, the combination of components at the nanoscale level with joint plastic deformation under pressure, ensuring the dislocation mass transfer, which is deformation mixing at the molecular level. In other words, there is a mutual distribution components (CC and SHWC) to level the molecular or cluster, the preservation of the native structure, in particular for gold remains the oxidation state (+3),

2) at a temperature of from -5°C to 50°C, a pressure of from 50 to 100 MPa and duration of response to 5 minutes of simultaneous chemical interaction of the initial reagents, namely SHVC or gold salts, recovered in the course of the synthesis to nanoscale gold(0, +2, +3), on the one hand, with the hydroxyl groups of salt (salt) group with the formation of esters of the General Ledger and on the other hand - with carboxyl and hydroxyl groups of citric acid with the formation of a stable chelate polycomplexes citric acid-gold-Ledger lemon-gold, CC gold, and mixtures thereof. Such polycomplexes stability in General is not inferior to the covalently bound systems, as they contain recurring polychelate fragments distributed on macrocopy of macrocomplex;

3) at a temperature from 50°C to 125°C, a pressure of from 200 to 1000 MPa and duration of the reaction up to 30 minutes, is carried out simultaneous chemical interaction of hyaluronic acid with citric acid in a free form, where the alcohol group of hyaluronic acid at the 6th carbon atom of N-acetyl-glucosamine tarifitsirovana hydroxyl group of citric education Sobolev-glucuronic acid 6-O-citrol chitin), and where the carboxyl group of D-glucur the new balance of hyaluronic acid etherification hydroxyl group of citric acid with the formation of CC-O-acyl citric acid: GK-LK-gold, GK-gold, LK-gold and mixtures thereof. When this process goes on the mechanism of nucleophilic substitution. As the acid catalyst is a Lewis acid - SHUK or other gold-bearing acid.

Thus, the composition of the nanocomposites nanocomposite changes as follows: (1) prevails ZHUK, distributed in the General Ledger at the molecular level, (2) is dominated by the chelates of the main component - LK-gold-Ledger, and (3) - chelates CC-O-acyl citric acid - HK-LK-gold.

At the last stage in the reactor if necessary add the estimated amount of the stabilizer (cysteine or glutathione), which forms a covalent bond with the previously stable gold, and ensures durability bioactive nanocomposites nanocomposite as a whole.

The scheme of reactions:

I) the Interaction of GC with LK in terms of targetalert (tverdofaznoi) and acid catalysis with the formation of the predominant products a and B.

The product of (A) poly(D-glucuronic acid 6-O-citrol chitin) or poly[2-(acetylamino)-2-deoxy-6-O-{6-[1,2-dicarboxy-1-(carboxymethyl)ethyl]-3-O-β-D-glucopyranosyl}-α-D-glucopyranose].

The product (B) CC-O-acyl citric acid or poly[2-(acetylamino)-2-deoxy-3-O-{6-[1,2-dicarboxy-1-(carboxymethyl)ethyl]-β-D-glucopyranose-ZIL}-α-D-glucopyranose].

II) the interaction of the products (a) and (B) in terms of targetalert with C the VK

III) the interaction of the products (a) and (B), chelating gold, targetalert with cysteine or glutathione.

or

IV) In the form of micelles target products are as follows: (for example, product A, LF gold and stabilizer - cysteine).

V) the main by-products of the interaction of GC, LC,SHVC, and also the introduction into the system of cysteine or glutathione in terms of targetalert (tverdofaznoi).

About the quantitative yield of the target products were judged according to FTIR spectral analysis of the initial reagents and reaction products. It is established that in the spectra of these products strip in the area 1150-1070 cm-1caused by asymmetric stretching vibrations of groups C-O-C, corresponds to the formation of simple ether - link copoly(D-glucuronic acid 6-O-citrol chitin). Bands in the area 1750-1735 cm-1connection GK-is-acyl citric acid. The nature of the bands in the region in 3430-3370 cm-1corresponding to the vibrations of-OH groups in the side 418, 441, 445 cm-1the corresponding fluctuations Au-O. the Presence of L-cysteine was determined by the following characteristic bands in the IR spectra: 1580-1590, 1600-1610 and 3000-3020 cm-1. For L-cysteine in the form of zwitterions (internal salt) characteristically show spectra of carboxylate ion (COO-, 1600 and 1390 cm-1), primary salts of amines, group NH3+(N-H, 3000-3500 cm-1), sulfhydryl groups (-SH, 2550 cm-1). Interaction (S-Au) is confirmed by the decrease (-SH, 2546 cm-1or the disappearance of the band (-SH, 2550 cm-1and the appearance of bands 1050 cm-1.

Glutathione is an amino acid that consists of residues, glutamic acid, cysteine and glycine, therefore, glutathione characterized by the same band, and cysteine. As for cysteine interaction (S-Au) is confirmed by the decrease (-SH, 2529 cm-1) or disappearance (-SH, 2529 cm-1and the appearance of bands 1079 cm-1.

The formation of LF gold was observed in the absorption spectra due to the well known phenomenon of surface plasmon resonance associated with the existence of surface electromagnetic waves at the boundary of metal and dielectric. The size of the resulting particles (nm) and their variation in size (share in %) were determined using transmission ELEH the throne microscope (FACT) and dynamic light scattering (DLS).

The invention can be illustrated by the following examples.

Example 1. The molar ratio of CC:citric acid = 1:1. Take 60 grams (15·10-2mole) of powdered sodium salt of ha (average molecular weight approximately 1000 kDa), 29 g (15·10-2mole) of citric acid and 10 g (2.5·10-2mole) soloconsolidation acid (SHVC), homogenizers in the mill at -18°C for 10-15 minutes Then the homogeneous powder mixture is fed into the power zone twin screw extruder where the material transporting capture elements and move it along the length of the cylinder during rotation. In the second and the third zone the material is subjected to shear strain due to the mixing elements consisting of Cams scored five pieces with the angle of rotation between the Cams 45°, 90° and 45° (reverse). The items are placed at different angles contributes to the formation of constipation in goods movement and as a consequence his best mixing and great physical stress.

The extruder has a measuring speed of rotation of the augers, the readings of which is proportional to the input voltage and load meter on augers, showing the value of the constant current drive.

The process is carried out with the automatic loading of material in a stream of nitrogen, the feed rate of the reaction mixture was maintained so the th, in order to maintain a given level of load on the augers. The speed of the augers were chosen in the range from 20 to 200 rpm·min-1. Load (current) without load 5 a, and solid-state reaction mixture is optimally supported 30-35 A. Temperature in the first zone of 5°C, in the second 5°C, in the third zone of 5°C. the Cycle is repeated 3 times. The duration of the process is ~10 minutes at a pressure of 50 MPa. The temperature for the 4th run is in the first zone 50°C, the second at 120°C, in the third zone of 120°C. the Cycle repeats 2 times. The duration of the process is ~10 minutes at a pressure of 200 MPa. On the 6th run, add the calculated amount of sodium hydroxide in the amount of 18.2 g, respectively, or on the basis of data of potentiometric titration of samples of the composite. Temperature on the 6th run is in the first zone of 5°C, in the second 5°C, in the third zone of 5°C. the Total process time ~20 minutes. The product yield is 116,0 g (99,0%). The degree of esterification of hydroxyl groups of ha for 6-carbon atom of N-acetyl-glucosamine Luke is 5%; carboxyl groups of D-glucoronidase balance 10%. The maximum absorption is 521 nm; the size of the gold particles corresponds to a value of 5.2 nm, the proportion of such particles 90%. The maximum absorption and the size of the gold particles remain unchanged during the year. The degree of filling of the composite gold is 4.3 wt %.

Example 2. Performed analogously to example 1, but in contrast, the molar ratio of CC:citric acid = 1:0.5 in. Then take 14.5 g (15·10-5mole) of citric acid and 5 g (12.5·10-3mole) soloconsolidation acid (SHWC). On the 6th run, add the calculated amount of sodium hydroxide in the amount of 9.1 g, respectively, or on the basis of data of potentiometric titration of samples of the composite.

The temperature for the 4th run is in the first zone 50°C, in the second 50°C, in the third zone 50°C. the Cycle repeats 2 times. Load (current) without load 5 a, and solid-state reaction mixture is optimally supported 10-15 A. the Duration of the process is -10 minutes at a pressure of 50 MPa. The product yield is 87,7 g (99,0%). The degree of esterification of hydroxyl groups of ha for 6-carbon atom of N-acetyl-glucosamine LK is less than 5%; carboxyl groups of D-glucoronidase residue less than 5%. The maximum absorption is 523 nm; the size of the gold particles corresponds to a value of 12.0 nm, the proportion of such particles 67%. The maximum absorption and the size of the gold particles remain unchanged during the year. The degree of filling of the composite gold is 2.9 wt. %.

Example 3. Performed analogously to example 1, but in contrast, the molar ratio of CC:citric acid = 1:0,01. Then take 0.29 grams (15·10-4mole) lemon sour is s and 1·10 -1g (2.5·10-4mole) soloconsolidation acid (SHWC). On the 6th run, add the calculated amount of sodium hydroxide in the amount of 18.2·10-2g, respectively, or on the basis of data of potentiometric titration of samples of the composite.

The temperature for the 4th run is in the first zone 50°C, in the second 50°C, in the third zone 50°C. the Cycle repeats 2 times. Load (current) without load 5 a, and solid-state reaction mixture is optimally supported 20-25 A. the Duration of the process is ~10 minutes at a pressure of 100 MPa. The degree of esterification of hydroxyl groups of ha for 6-carbon atom of N-acetyl-glucosamine Luke is 5%; carboxyl groups of D-glucoronidase balance 10%. The maximum absorption is 517 nm; the size of the gold particles corresponds to a value of 1.1 nm, the proportion of such particles 87%. The maximum absorption and the size of the gold particles remain unchanged during the year. The degree of filling of the composite gold is 8.3·10-2wt. %.

Example 4. Performed analogously to example 1, however in contrast instead of citric acid take sodium citrate. The molar ratio of CC:citrate = 1:2. Then take 77,22 g (3·10-1mole) of sodium citrate and 19,31 g (4,9·10-2mole) soloconsolidation acid (SHWC). On the 6th run of sodium hydroxide is not added. Load 30-35A, pressure 140 M the and. The total process time ~20 minutes. The product yield is 154,96 g (99,0%). The degree of esterification of hydroxyl groups of ha for 6-carbon atom of N-acetyl-glucosamine LK is less than 5%; carboxyl groups of D-glucoronidase balance of 6%. The maximum absorption at 525 nm; the size of the gold particles corresponds to a value of 15.0 nm, the proportion of such particles 97%. The maximum absorption and the size of the gold particles remain unchanged during the year. The degree of filling of the composite gold is 6.2 wt. %.

Example 5. Performed analogously to example 1, however in contrast instead of citric acid take magnesium citrate, and instead SHVC solutocapillary potassium. The molar ratio of CC:magnesium citrate = 1:1. Ledger charge 100 g (25·10-2mol), magnesium citrate 53.47 USD g (25·10-2mole) and 26,74 g (3,4·10-2mole) zolotoizvlekatelnoj potassium. On the 6th run of sodium hydroxide is not added. Load 40-45 And a pressure of 180 MPa. The total process time ~20 minutes. The product yield is 178,41 g (99,0%). The degree of esterification of hydroxyl groups of ha for 6-carbon atom of N-acetyl-glucosamine LK is less than 5%; carboxyl groups of D-glucoronidase balance of 6%. The maximum absorption is 519 nm; the size of the gold particles corresponds to a value of 3.0 nm, the proportion of such particles 72%. The degree of filling of the composite gold is 3.75 wt. %.

Example 6. Performed analogously to example 1, but in contrast to the him instead of the sodium salt of the Ledger taken a mixed sodium-calcium salt at a molar ratio of sodium:calcium = 2:1. The product yield is 116,0 g (99,0%). The degree of esterification of hydroxyl groups of ha for 6-carbon atom of N-acetyl-glucosamine Luke is 5%; carboxyl groups of D-glucoronidase balance 10%. The maximum absorption is 520 nm; the size of the gold particles corresponds to a value of 5.0 nm, the proportion of such particles 91%. The maximum absorption and the size of the gold particles remain unchanged during the year. The degree of filling of the composite gold is 4.3 wt. %.

Example 7. Performed analogously to example 1, however in contrast instead of the sodium salt of the Ledger taken mixed sodium-zinc salt at a molar ratio of sodium:zinc = 2:1. The product yield is 116,0 g (99,0%). The degree of esterification of hydroxyl groups of ha for 6-carbon atom of N-acetyl-glucosamine Luke is 5%; carboxyl groups of D-glucoronidase balance 10%. The maximum absorption is 520 nm; the size of the gold particles corresponds to a value of 5.1 nm, the proportion of such particles 90%. The maximum absorption and the size of the gold particles remain unchanged during the year. The degree of filling of the composite gold is 4.3 wt. %

Example 8. Performed analogously to example 1, however in contrast instead of the sodium salt of the Ledger taken mixed sodium-gold salt at a molar ratio of sodium:copper = 2:1. On the 7th run, add the calculated amount of glutathione in the number 46,0 g (1.5·10-1supposedly is). The temperature regime at the 7th run is in the first zone 20°C, in the second 20°C, in the third zone of 20°C. the Total process time ~25 minutes. The product yield is 161.6 g (99,0%). The degree of esterification of hydroxyl groups of ha for 6-carbon atom of N-acetyl-glucosamine Luke is 5%; carboxyl groups of D-glucoronidase balance 10%. The maximum absorption is 518 nm; the size of the gold particles corresponds to a value of 2.4 nm, the proportion of such particles 83%. The maximum absorption and the size of the gold particles remain unchanged during the year. The degree of filling of the composite gold is 3.1 wt. %.

Example 9. Performed analogously to example 1, however in contrast instead of the sodium salt of the Ledger taken mixed salt Ledger:CMC at a molar = 1:20 in the amount of 60 g At the 7th run, add the calculated amount of cysteine in the number of 18.1 g (1.5·10-1mole). The temperature regime at the 7th run is in the first zone 20°C, in the second 20°C, in the third zone of 20°C. the Total process time ~25 minutes. The product yield is 134,0 g (99,0%). The degree of esterification of hydroxyl groups of ha for 6-carbon atom of N-acetyl-glucosamine Luke is 5%; carboxyl groups of D-glucoronidase balance 10%. The maximum absorption is 519 nm; the size of the gold particles corresponds to a value of 2.9 nm, the proportion of such particles 81%. The maximum absorption and the size of the gold particles remains unchanged during the year is. The degree of filling of the composite gold is 3.7 wt. %.

Example 10. Performed analogously to example 1, but unlike him take of 300.0 g (75·10-2mole) of powdered sodium salt SC, 100.0 g (5.2 x 10-1mole) of citric acid, 25,0 g (10,5·10-1mole) of sodium salt of CMC, 25,0 g of sodium salt of the GOC (5,5·10-3mole), homogenizers in the mill at 20°C for 10-15 min Speed of rotation of the augers in the range from 20 to 100 rpm·min-1. Load (current) without load 5 a, and solid-state reaction mixture is optimally supported 40-45 A. the Temperature in the first, second and third zone 50°C. the Cycle is repeated 3 times. The temperature for the 4th run is in the first zone -15°C, in the second -15°C, in the third zone -15°C. On the 4th run to the reaction mixture is added DMSO to 10% by weight of the composition and dosed solutocapillary potassium (K[AuI4]·nH2O, n=3 or 4) in an amount of 100.0 g (125·10-3mole). The cycle is repeated 4 times. The temperature regime in the next two cycles is in the first area 105°C, the second 110°C, in the third zone of -15°C. the Total duration of the process is ~30 minutes at a pressure of 400 MPa. The product yield is 544,5 g (-100,0%). The degree of esterification of hydroxyl groups of ha for 6-carbon atom of N-acetyl-glucosamine LK 10%; carboxyl groups of D-glucoronidase balance of 22%. Max is ogloszenia is 521 nm; the size of the gold particles corresponds to a value of 6.0 nm, the proportion of such particles is 98%. The maximum absorption and the size of the gold particles remain unchanged during the year. The degree of filling of the composite gold is 4.6 wt. %.

Example 11. Performed analogously to example 1, but unlike him take of 300.0 g (75·10-2mole) of powdered sodium salt SC, 100.0 g (5.2 x 10-1mole) of citric acid, homogenizers in the mill at 20°C for 10-15 min Speed of rotation of the augers in the range from about 20 to 80·min-1. Load (current) without load 5 a, and solid-state reaction mixture is optimally supported 40-45 A. the Temperature in the first, second and third zone 50°C. the Cycle is repeated 3 times. The temperature for the 4th run is in the first zone -15°C, in the second -15°C, in the third zone -15°C. On the 4th run dosed solutocapillary potassium (K[AuI4]·nH2O, n=3 or 4) in an amount of 100.0 g (125·10-3mole). The cycle is repeated 10 times. The temperature regime in the next five cycles is in the first zone 110°C, in the second 125°C, in the third zone -15°C. For 16-m run, add the calculated amount of glutathione in the amount of 30.0 g (9,8·10-2mole). Temperature on the 16th run is in the first zone 20°C, in the second 20°C, in the third zone of 20°C. the Cycle repeats 2 times. The total duration of the process is ~60 minutes the ri pressure of 600 MPa. The product yield is 544,5 g (~100,0%). The degree of esterification of hydroxyl groups of ha for 6-carbon atom of N-acetyl-glucosamine Luke is 13%; carboxyl groups of D-glucoronidase balance 30%. The maximum absorption is 522 nm; the size of the gold particles corresponds to a value of 8.0 nm, the proportion of such particles 97%. The maximum absorption and the size of the gold particles remain unchanged during the year. The degree of filling of the composite gold is 4.6 wt. %.

Example 12. to 120.0 mg (3·10-4mole) of powdered sodium salt of hyaluronic acid, 20.0 mg (1·10-4mole) of citric acid and 4.0 mg (1·10-5mole) soloconsolidation acid (SHVC) homogenized mill at -10°C for 10-15 minutes Then the homogeneous powder mixture is placed on the lower anvil Bridgman (the diameter of the working surface =3 cm), cover the top of the anvil, the anvil put under a press and subjected to pressure of 200 MPa at 20°C when the angle of rotation of the lower anvil 250° during 1 min. Then relieve pressure, remove the anvil from the press. The product yield is 141,1 mg (98,0%). The degree of esterification of hydroxyl groups of ha for 6-carbon atom of N-acetyl-glucosamine Luke is 5%; carboxyl groups of D-glucoronidase balance 10%. The maximum absorption is not specified; the size of the gold particles correspond to a value of 1.8 nm, the proportion of such particles 88%. Maximum pogloshena and the size of the gold particles remain unchanged during the year. The degree of filling of the composite gold is 1.4 wt. %.

Example 13. to 120.0 mg (3·10-4mole) of powdered sodium salt SC, 120,0 mg (6,2·10-4mole) of citric acid and 60.0 mg (7,5·10-5mole) zolotoizvlekatelnoj potassium (K[AuI4]·nH2O, n=3 or 4) homogenize in the mill at 5°C for 10-15 minutes Then the homogeneous powder mixture is placed on the lower anvil Bridgman (the diameter of the working surface = 3 cm), cover the top of the anvil, the anvil put under a press and subjected to pressure of 450 MPa at 90°when the angle of rotation of the lower anvil 200° within 30 seconds. Further relieve pressure, remove the anvil from the press. The product yield is 294,0 mg (98,0%). The degree of esterification of hydroxyl groups of ha for 6-carbon atom of N-acetyl-glucosamine Luke is 11%; carboxyl groups of D-glucoronidase balance of 27%. The maximum absorption is 527 nm; the size of the gold particles corresponds to a value of 20.0 nm, the proportion of such particles is 95%. The maximum absorption and the size of the gold particles remain unchanged during the year. The degree of filling of the composite gold is 5.1 wt. %.

Example 14. Performed analogously to example 12, however, in contrast to initially deform the Ledger and W at a pressure of 300 MPa, 50°C, the angle of rotation of the lower anvil 250° for 1 minute Then take off the pressure, and add 4.0 mg (1·10-5mole) gold is hloristovodorodnykh acid (SHWC). Then the homogeneous powder mixture is placed on the lower anvil Bridgman (the diameter of the working surface = 3 cm), cover the top of the anvil, the anvil put under a press and subjected to pressure of 50 MPa at 20°C when the angle of rotation of the lower anvil 200° for 30 sec. Then raise the pressure up to 1000 MPa at 120°C when the angle of rotation of the lower anvil 350° within 2 minutes. The product yield is 136,8 mg (95%). The degree of esterification of hydroxyl groups of ha for 6-carbon atom of N-acetyl-glucosamine LK is 20%; carboxyl groups of D-glucoronidase balance 50%. The maximum absorption is not specified; the size of the gold particles corresponds to a value of 1.3 nm, the proportion of such particles is 85%. The maximum absorption and the size of the gold particles remain unchanged during the year. The degree of filling of the composite gold is 1.46 wt. %.

Example 15. Performed analogously to example 12, however in contrast instead of the sodium salt of the Ledger taken a mixed sodium-calcium salt at a molar ratio of sodium:calcium = 2:1. The product yield is 141,1 mg (98,0%). The degree of esterification of hydroxyl groups of ha for 6-carbon atom of N-acetyl-glucosamine LK 10%; carboxyl groups of D-glucoronidase balance of 24%. The degree of esterification of hydroxyl groups of ha for 6-carbon atom of N-acetyl-glucosamine Luke is 5%; carboxyl groups of D-glucurono the th balance of 10%. The maximum absorption is not specified; the size of the gold particles correspond to a value of 1.8 nm, the proportion of such particles is 85%. The maximum absorption and the size of the gold particles remain unchanged during the year. The degree of filling of the composite gold is 1.4 wt. %.

Example 16. Performed analogously to example 13, however in contrast instead of the sodium salt of the Ledger taken mixed sodium-aluminum salt at a molar ratio of sodium:aluminum = 3:1. The product yield is 294,0 mg (98,0%). The degree of esterification of hydroxyl groups of ha for 6-carbon atom of N-acetyl-glucosamine LK 10%; carboxyl groups of D-glucoronidase balance of 24%. The maximum absorption at 525 nm; the size of the gold particles corresponds to a value of 15.0 nm, the proportion of such particles is 95%. The maximum absorption and the size of the gold particles remain unchanged during the year. The degree of filling of the composite gold is 5.1 wt. %.

Example 17. Performed analogously to example 12, but unlike him after removal of the pressure type of 9.4 mg (3,0·10-5mole) glutathione. Then the homogeneous powder mixture is placed on the lower anvil Bridgman (the diameter of the working surface = 3 cm), cover the top of the anvil, the anvil put under a press and subjected to pressure of 30 MPa at 20°C when the angle of rotation of the lower anvil 200° within 1 min. Yield 148.8 mg (97,0%). the degree of esterification of hydroxyl groups of ha for 6-carbon atom of N-acetyl-glucosamine Luke is 5%; carboxyl groups of D-glucoronidase balance 10%. The maximum absorption is 522 nm; the size of the gold particles corresponds to a value of 10.0 nm, the proportion of such particles 92%. The maximum absorption and the size of the gold particles remain unchanged during the year. The degree of filling of the composite gold is 1.34 wt. %.

Example 18. Performed analogously to example 13, but unlike him after removal of the pressure type 27.3 mg (9,7·10-6mole) of cysteine. Then the homogeneous powder mixture is placed on the lower anvil Bridgman (the diameter of the working surface =3 cm), cover the top of the anvil, the anvil put under a press and subjected to pressure of 50 MPa at 20°C when the angle of rotation of the lower anvil 200° within 1 min. Yield is 320,8 mg (98,0%). The degree of esterification of hydroxyl groups of ha for 6-carbon atom of N-acetyl-glucosamine LK 10%; carboxyl groups of D-glucoronidase balance of 22%. The maximum absorption at 525 nm; the size of the gold particles corresponds to a value of 15.0 nm, the proportion of such particles 94%. The maximum absorption and the size of the gold particles remain unchanged during the year. The degree of filling of the composite gold is to 4.68 wt. %.

The examples clearly show that a versatile, environmentally safe way to obtain a water-soluble bioactive nanocomposite is, includes a modified citric acid or a salt of citric acid salt of hyaluronic acid as a matrix and the gold nanoparticles as a filler, in one technological mode in the absence of a liquid medium to obtain the desired products in high yields. The method does not require large energy, labor and vadastra, allows it to be used as the source of a variety of reagents, including water-insoluble salt of the code.

Achieved a synergistic combination of properties of components contained in the composition of the bioactive nanocomposites nanocomposite, in particular, strengthening antibacterial and antioxidant properties, along with toxicity. Achieved high stability LF gold in solid form, and when dissolved in water, keeping accurately known concentration of the main components. By changing the ratio of components, bioactive nanocomposites nanocomposite, gold compounds function as prodrugs, creating a way to "delivery" gold to areas of inflammation. All this gives the potential implementation of the principle of ERT, photothermal therapy, photo - and radiosensibility, chemotherapy, treatment of rheumatoid arthritis, antovic therapy.

For the purposes of aesthetic dermatology and plastic surgery for intradermal promotes the proliferation of fibroblasts and neoangiogenesis. I.e. stimulates about the establishment of new cells improving skin elasticity and reducing wrinkles.

1. The method of obtaining water-soluble bioactive nanocomposites nanocomposite comprising a modified citric acid or a salt of citric acid salt of hyaluronic acid as a matrix and the gold nanoparticles as a filler, characterized by the fact that carry out chemical interaction of solid-phase powders of salts of hyaluronic acid, citric acid or salts of citric acid and soloconsolidation acid or gold salts at temperatures from -18° to 125°C, under simultaneous pressure from 50 to 1000 MPa and shear strain in mechanochemical reactor.

2. The method according to p. 1, characterized in that the hyaluronic acid salt using salt selected from the range: tetraalkylammonium, lithium, sodium, potassium, calcium, magnesium, barium, zinc, aluminum, copper, gold or mixed salt of hyaluronic acid of the above number or Hydrosol hyaluronic acid.

3. The method according to p. 2, characterized in that the salt of hyaluronic acid is a sodium salt or a mixed salt, or hydrometridae salt.

4. The method according to p. 1, characterized in that as salts of citric acid using sodium citrate or potassium citrate, or magnesium citrate.

5. The method according to p. 1, characterizing the I, as gold salts used solutocapillary potassium, tetrachloroaurate (III) potassium tetrachloroaurate (III) ammonium, tetrachloroaurate (III) sodium, tetrabromophenol (III) sodium, dicyanoaurate potassium, tetracenomycin (III) potassium, disulfiram (I) ammonium bis(thiosulfate)Aurat (I) sodium, chloro(triphenylphosphine)gold (I).

6. The method according to p. 1, characterized in that additionally introduced into the reactor stabilizer.

7. The method according to p. 6, characterized in that the stabilizer is used as glutathione or cysteine.

8. The method according to p. 1, characterized in that the molar ratio of the salt of hyaluronic acid to citric acid is from 1:0.01 to 1:20, respectively.

9. The method according to p. 1, characterized in that the molar ratio of soloconsolidation acid to citric acid is in the range from 1:1 to 1:20, respectively.

10. The method according to p. 1, characterized in that the molar ratio of zolotoizvlekatelnoj potassium to citric acid is in the range from 1:1 to 1:20, respectively.

11. The method according to p. 7, characterized in that the molar ratio of soloconsolidation acid for glutathione is in the range from 1:1 to 1:15, respectively.

12. The method according to p. 7, characterized in that the molar ratio of soloconsolidation acid to cysteine is in a pre is Elah from 1:1 to 1:20, respectively.

13. The method according to p. 7, characterized in that the molar ratio of zolotoizvlekatelnoj potassium to the glutathione is in the range from 1:1 to 1:15, respectively.

14. The method according to p. 7, characterized in that the molar ratio of zolotoizvlekatelnoj potassium to the cysteine is in the range from 1:1 to 1:20, respectively.

15. The method according to p. 1, characterized in that the duration of the effects of pressure and shear strain is in the range from 0.1 to 60 minutes, in particular 6 minutes at a pressure of 1000 MPa.

16. The method according to p. 1, characterized in that as a mechanochemical reactor can be used, in particular, Bridgman anvils.

17. The method according to p. 1, characterized in that the mechanochemical reactor is a machine screw of the type selected from the set: twin screw extruder with a unidirectional rotation of the screw, twin-screw extruder with oppositely directed rotation of the screw, twin-screw extruder with a set of Cams of different types: transport, locking, peretiraya.

18. The method according to p. 1, characterized in that further added, at least one stabilizing additive.

19. The method according to p. 18, characterized in that as a stabilizing additive used substance selected from the group of carboxymethyl cellulose (CMC), tetraalkylammonium, lithium is, sodium, potassium, calcium, magnesium, barium, zinc, aluminum, copper, gold or mixed salt of CMC from the above number or Hydrosol CMC, hydroxyethyl cellulose, hydroxypropylcellulose.

20. The method according to p. 1, characterized in that the molar ratio of the salt of hyaluronic acid for stabilizing additive in the range from 100:1 to 1:1.



 

Same patents:

FIELD: medicine, pharmaceutics.

SUBSTANCE: invention refers to chitosan carboxyalkylamide hydrogel and may be used for cosmetic and dermatological treatment of skin burns. Chitosan carboxyalkylamide hydrogel of pH close to that of skin and making 6.5 to 7.2 contains 40 to 90 mole % of the groups of N-carboxyalkylaminde D-glucosamine of formula (I) wherein n represents an integer 1 to 8, 60 to 10 mole % of the protic groups of D-glucosamine, and 5 to 15 mole % of the groups of N-acetyl-D-glucosamine. A method for preparing said hydrogel involves preparing an acid solution of chitosan of a degree of acetylation of 85 to 95%, providing a reaction of produced additive chitosan salt in an aqueous solution of diorganic acid and correcting pH of the prepared solution.

EFFECT: preparing the anhydrous product of chitosan carboxyalkylamide prepared by hydrogel dehydration.

12 cl, 1 dwg

FIELD: chemistry.

SUBSTANCE: biocompatible, biodegradable porous composite material contains chitosan and hydrosilicate filler in amount of 0.05-10% of the weight of chitosan and has a system of through pores with size of 5-1000 mcm. The method of producing the material involves mixing hydrosilicate filler, which is pre-dispersed in an aqueous medium with pH=5-7 in an ultrasonic field with frequency v=20-100 kHz for 5-60 minutes, with chitosan in an amount which corresponds to its concentration in the solution of 1-4 wt %, the amount of the filler being equal to 0.05-10% of the weight of chitosan; the obtained mixture is then intensely mixed at temperature of 20-50°C for 20-60 minutes; concentrated acetic acid is added in an amount which enables to obtain, in the mixture of the aqueous solution, acetic acid with concentration of 1-3%; the mixture is intensely mixed at temperature of 20-50°C for 20-250 minutes and then cooled to temperature of -5 to -196°C; the solvent is removed in a vacuum; the obtained end material is treated with a neutralising agent, washed with water to pH=5-7 and then dried.

EFFECT: presence of a system of through pores and providing a stable porous structure of the material in aqueous medium, eliminating cytotoxicity.

6 cl, 9 ex, 6 dwg

FIELD: chemistry.

SUBSTANCE: biodegradable film contains pectin, chitosan, water, 1N hydrochloric acid, a plasticiser - glycerine and a structure-forming agent - 3% methyl cellulose solution.

EFFECT: obtaining a uniform biodegradable film without structural defects, which similar to household packaging polyethylene films on homogeneity, plasticity and strength.

2 dwg, 1 tbl

FIELD: chemistry.

SUBSTANCE: invention relates to synthetic polymer chemistry. The nanocomposite contains a matrix in form of a cross-linked salt of hyaluronic acid which is modified with sulphur-containing compounds and nanoparticles of a noble metal as filler. A film of the cross-linked salt of hyaluronic acid which is modified with sulphur-containing compounds is obtained through chemical reaction of the salt of hyaluronic acid with a mixture of two sulphur-containing compounds and with a cross-linking agent, under conditions with pressure between 50 and 300 MPa and shear deformation in a mechanical reactor at temperature between 20 and 30°C. The reactor used to obtain the film is a Bridgman anvil.

EFFECT: invention enables to obtain a range of new bioactive nanocomposites with quantitative output and in the absence of a liquid medium, where the method does not require high energy, labour and water consumption and significantly increases efficiency of the composite; in particular, resistance to decomposition in the presence of hydroxyl radicals is 2-3 times higher compared to the control result.

16 cl, 7 ex

FIELD: medicine.

SUBSTANCE: solution of viable breast cells after enzymatic degradation in a collagenase solution of analysed tissue at temperature 37°C for 30-35 minutes, are applied on a biomaterial of a native form of hyaluronic acid, stored at room temperature and constant humidity 30-50 %, and cell viability is controlled by discoloration of cell monolayer.

EFFECT: invention allows producing a viable cell monolayer and providing their vital activity for 2-3 hours.

1 tbl, 2 dwg

FIELD: chemistry.

SUBSTANCE: articles based on biologically active polymer materials can be used in surgery when treating wounds and as materials for temporary replacement of body tissue, in biotechnology for obtaining matrices for growing cell cultures, in pharmaceuticals as carriers of enzymes and other biologically active compounds. The cross-linking reagents are 2,4-derivatives of 3-oxa glutaric dialdehyde (2,2'-oxydiacetaldehyde). These compounds can be obtained via periodate oxidation of monosaccharides, nucleosides and nucleotides.

EFFECT: low toxicity and biocompatibility of chitosan-based materials is achieved by using cross-linking reagents.

2 dwg, 12 ex

FIELD: chemistry.

SUBSTANCE: invention relates to a method of obtaining polysaccharide fibre for making materials, specifically for making surgical suture materials absorbable in a human and a mammal body, absorbable and non-absorbable dressing material and absorbable textile matrix materials. The method is characterised by that 2.4-4.0 wt % solution of polysaccharide in dimethylacetamide which contains 4.56-10.00 wt % lithium chloride is mixed with 1.0-5.0 wt % poly-N-vinylpyrrolidone with molecular weight of 8-35 kDa or a metallopolymer complex - fine-grained silver stabilised by poly-N-vinylpyrrolidone in such an amount that, content of fine-grained silver relative polysaccharide dissolved in spinning solution ranges from 0.07 to 0.87 wt %. Mass ratio of polysaccharide: metallopolymer complex equals 88.0-99.0:1.0-12 wt %. The mixture is intensely stirred, held, filtered, degassed and the obtained spinning solution is extruded at room temperature into an alcohol deposition tank in form of water-soluble aliphatic C2 and C3 alcohols. The fibre is then processed in plastification and washing tanks and dried.

EFFECT: obtaining fibre with good deformation and strength properties.

1 dwg, 4 tbl, 29 ex

FIELD: chemistry.

SUBSTANCE: invention relates to taxane, especially to paclitaxel and docetaxel covalently bonded with hyaluronic acid or a hyaluronic acid derivative, used as active substance in pharmaceutical compositions which are used in oncology, for treating tumours, autoimmune diseases and restenosis, as well as a coating for stents and medical devices. The covalent bond is formed between hydroxyl groups of taxane and carboxyl groups or hydroxyl groups of hyaluronic acid or hyaluronic acid derivatives, or amino groups of deacetylated hyaluronic acid. Bonding can take place using a linking compound (spacer compound) which bonds taxane with hyaluronic acid or hyaluronic acid derivative under the condition that, the linking compound is different from hydrazide.

EFFECT: proposed taxane has high therapeutic effectiveness when treating oncological diseases, autoimmune diseases and restenosis, dissolves in water without reduction of its pharmacological activity and is not toxic, which leads to overcoming hypersensitivity and anaphylaxis.

46 cl, 20 ex, 4 dwg

FIELD: medicine.

SUBSTANCE: composition includes chitosan gel, which has bactericidal properties, representing matrix for including in it of water solution for "Adgelon" injection, which contains serum glycoprotein from blood of livestock, possessing biological activity in supersmall doses 10-9-10-15 mg/ml, and calcium salt - preparation "Adgelon". In obtaining composition medicine "Adgelon" is immobilised on chitosan gel.

EFFECT: composition provides increased ability to stimulate proliferation of osteoblasts, reparative processes of osteoformation with recovery by means of morphologically normal bone matrix.

2 cl, 3 ex

FIELD: chemistry, medicine.

SUBSTANCE: complex matrix consists of at least one biologically compatible polymer of natural origin, structured with sewing agent, which represents two- or multi-functional molecule, selected from epoxides, epihalohydrines and divinyl sulphone, on said polymer inoculated are chains with molecular weight less than 50000 daltons, selected from polymers of natural origin of small size, preferably, derivatives of cellulose or other biological polymer derivatives which naturally are not present in human organism, and/or non-polymerised chains with properties of oxidation inhibitors or ability to inhibit reactions of matrix decomposition, preferably, vitamins, enzymes or molecules, consisting of one or several cycles, degree of inoculation, expressed as ratio of moles of inoculated molecules and quantity of moles of polymer units, constitutes from 10 to 40%. Also described are method of obtaining such matrix and its application for separation, replacement, filling or addition of biological fluid or tissues.

EFFECT: increase of application efficiency.

20 cl, 7 ex, 2 tbl, 1 dwg

FIELD: chemistry.

SUBSTANCE: invention relates to natural polysaccharide polymers and can be used in medicine. The obtained water-soluble bioactive nanocomposite includes a melanin compound-modified hyaluronic acid salt as a matrix and gold nanoparticles as filler. The method includes chemical reaction of solid-phase hyaluronic acid powder, a melanin compound, aurichlorohydric acid or a gold salt in conditions of simultaneous pressure action in the range of 50 to 1000 MPa and shearing deformation in a mechanochemical reactor at temperature of -18° to 110°C.

EFFECT: invention enables to obtain a water-soluble bioactive nanocomposite with high output of the end product and high content of gold.

4 cl, 18 ex

FIELD: chemistry.

SUBSTANCE: stabiliser includes modified chitosan which is obtained by modifying chitosan particles located in an emulsion of an organic solvent - water, with pH 6.0-6.5, by first reacting a mixture consisting of a carboxylic acid in an organic solvent and a condensing agent, and then with an organic base, wherein the carboxylic acid used is either palmitic acid or stearic acid or dodecanoic acid, the condensing agent used is a mixture of hydroxysuccinimide and an aliphatic carbodiimide or formaldehyde and an aliphatic isocyanide, and the organic base used is triethylamine.

EFFECT: effective liposome composition stabiliser which can be obtained using a simple method.

8 cl, 3 tbl, 5 ex, 7 dwg

FIELD: chemistry.

SUBSTANCE: invention relates to the field of organic synthesis. A method of obtaining a water-insoluble sulphur-containing chitosan-based biopolymer includes interaction of chitosan with a thiomethylating agent, preliminarily obtained by saturation of a formaldehyde solution with gaseous H2S, with molar ratio chitosan: formaldehyde: hydrogen sulphide 1:6:4, at a temperature of 60°C for 20-25 hours.

EFFECT: invention ensures obtaining the water-insoluble sulphur-containing chitosan-based biopolymer, which possesses a complexing activity to ions of noble metals (Pd, Pt).

1 ex, 1 tbl

FIELD: biotechnologies.

SUBSTANCE: skins of pond fish are flushed with cold flushing water during 10-15 minutes. They are crushed to the size of 2-3 mm. Water extraction is performed at the temperature of 40-45°C during 40-50 minutes at the ratio of crushed skins to water, which is equal to 1:1 at periodic mixing. Then, they are filtered; liquid fraction is dried in a spraying drier at the drier outlet product temperature of 60-65°C during 15-25 minutes so that hyaluronic acid is obtained. Solid fraction is subject to bleaching during 12 hours with hydrogen peroxide-salt solution that is prepared by mixing of 1 l of 3% hydrogen peroxide and 20 g of sodium chloride. Treatment of bleached solid fraction is performed with 1.0-1.2% solution of sodium hydroxide during 24 hours at the temperature of 20-25°C with further neutralisation of the obtained mixture with 3% boric acid solution. Treatment of swollen solid fraction is performed with Pancreatin ferment preparation solution taken in the quantity of 0.5-0.6% to the weight of solid fraction during 1.5-2.0 hours at the temperature of 37-40°C. Flushing of solid fraction is performed with cold flushing water for removal of Pancreatin residues so that collagen is obtained. The obtained collagen, depending on the purpose, is supplied for drying in drying chambers with forced air circulation at the temperature of 18-20°C during 12 hours and storage in dry ventilated rooms at the temperature of not higher than 20°C during 24 months or frozen to the temperature of minus 18 - minus 20°C and stored at the temperature of minus 18 - minus 20°C during 24 months. The liquid fraction dried in the spraying chamber is stored at the temperature of 0-4°C during 12 months or dissolved in physiological buffer solution.

EFFECT: improvement of the method.

2 dwg, 1 tbl, 1 ex

FIELD: biotechnologies.

SUBSTANCE: production method of glucan-chitosan complex from yeast biomass of brewing waste involves mechanical and ultrasonic treatment of yeast biomass, destruction of proteins by treatment of the obtained suspension using alkali reagents with further extraction of a target product. As biomass, Saccharomyces living yeast is used. First, yeast is frozen to -15°C during 24 hours. After mechanical destruction, biomass is treated for 15 minutes at 20°C in an ultrasonic bath with frequency of an emitter of 35 kHz and voltage of 285 W. Biomass is acidified with chlorhydric acid till pH=5.5 and treated with ferment preparation in the amount of one pellet containing lipase - 3500 units of Ph.Eur., amylase - 4200 units of Ph.Eur. and protease - 250 units of Ph.Eur. per kilogramme of biomass in terms of dry substance; then, lipid components of yeast are removed. Fermentation is performed at t=20-29°C during 30-60 minutes. Destruction of proteins is performed at 55°C by means of a water bath during 60 minutes by treatment using 4% water solution of caustic soda at the ratio of yeast biomass and alkali, which is equal to 1:4. The medium is neutralised and hydrosol of glucan-chitosan complex is deposited by centrifugation during 10 minutes. The deposit is dried at t=55°C during 48 hours.

EFFECT: invention allows improving the quality of the obtained complex and its biological activity.

3 ex

FIELD: chemistry.

SUBSTANCE: invention relates to production of hydroxyalkyl derivatives of polysaccharides. The method of producing 2,3-dihydroxypropyl chitosan involves reacting chitosan with glycidol in the presence of hydrochloric acid with ratio glycidol:chitosan:hydrochloric acid = (2-6):1:1, at room temperature until a gel forms. The mixture is then heated at 55-65°C for 12-14 hours and the reaction mass is treated with water. The mixture is then deposited, subjected to hot extraction with water-soluble alcohols or ketones and dried.

EFFECT: invention simplifies the method of production and output of the end product and improves sorption properties of the compound.

1 tbl, 3 ex

FIELD: medicine, pharmaceutics.

SUBSTANCE: there are presented: using benzophenanthridine alkaloid salts for preparing therapeutic agents for treating tumours, wherein the alkaloid salt is found in the form luteic, phosphatidic or hyaluronic acid, the benzophenanthridine alkaloid salt with phosphatidic acid or hyaluronic acid, and a based pharmaceutical composition for treating tumours.

EFFECT: what is shown is cytotoxic activity of the sanguinarine salts according to the invention at least twice increased in all studied tumour cell lines in relation to the chloride salt; it is suggested to be caused by higher absorption by the tumour cells.

12 cl, 8 ex

FIELD: medicine, pharmaceutics.

SUBSTANCE: invention refers to a recovered imidised biologically compatible polymer functionalised by an imide group. The above polymer is selected from the group consisting of polyethylene oxide, partially or completely hydrolysed by polyvinyl alcohol, polyvinylpyrrolidone, polyethyloxazoline, polyoxypropylene oxide block copolymers (poloxamers and meroxapol), polyethylene oxide and poloxamine copolymer, carboxymethyl cellulose and hydroxyalkylated cellulose, polypeptides, polysaccharides, carbohydrates, polysaccharose, hyaluronic acid, dextran, heparin sulphate, keratan sulphate, chondroitin sulphate, heparin, alginate, gelatin, collagen, albumin, ovalbumin, complex polyphosphoesters, polylactides, polyglycolides, polycaprolactones, polyamides, polyurethanes, polyesteramides, polyorthoesters, polydioxanones, polyacetals, polyketals, polycarbonates, polyorthocarbonates, polyphosphazenes, polyhydroxybutyrates, polyhydroxyvalerates, polyalkylene oxalates, polyalkylene succinates, polymaleic acids, polyamino acids, polyvinyl alcohol, polyvinylpyrrolidone, polyhydroxy cellulose, chitin, chitosan, and copolymers, ternary copolymers, or combinations or mixtures of the aforementioned materials. Also, the invention refers to a composition for a tissue adhesive, a medical device and a pharmaceutical composition.

EFFECT: invention represents additionally modified or functionalised imidised polymers.

9 cl, 2 ex, 20 dwg

FIELD: biotechnologies.

SUBSTANCE: fish roe is homogenised. Fish roe hydrolysis is carried out with a ferment preparation "Collagenase" in presence of an inhibitor for 10-12 hours. Chitosan is added to the produced hydrolysate at the ratio of 0.5-1.0:1.0. Components are mixed.

EFFECT: invention makes it possible to accelerate process of chitosan-nucleic complex production.

1 dwg, 1 tbl, 3 ex

FIELD: biotechnologies.

SUBSTANCE: method includes depolymerisation of a high-molecular chitosan with hydrogen peroxide. The process of chitosan depolymerisation is carried out in a double-phase system. The solid phase is activated chitosan with Mav = 450-650 kDa and the average particle size of 0.05-0.20 mm. The liquid phase is a water solution of H2O2 with concentration of H2O2 in a reaction system equal to 1-7%. The reaction is carried out for 120-180 minutes at 70°C. Then phase separation of produced chitosan homologs is carried out by means of filtration via paper or textile surface of the produced reaction mixture. The produced filtrate contains water-soluble chitosan oligomers.

EFFECT: invention makes it possible to quantitatively control extent of conversion of an initial high-molecular chitosam into oligomer and low-molecular structures of its homologs.

1 tbl, 1 ex

FIELD: electricity.

SUBSTANCE: method for manufacture of powerful SHF transistor includes application of a solder layer to the flange, shaping of pedestal, application of a sublayer fixing the transistor crystal to the pedestal, formation of p-type conductivity oriented at the plane (111) at the base substrate of single-crystalline silicon and auxiliary epitaxial layers, application of the basic layer and buffer layer for growing of epitaxial structure of a semiconductor device based on wide-gap III-nitrides, application of heat conductive layer of CVD polycrystalline diamond to the basic layer, removal of the basic substrate with auxiliary epitaxial layers up to the basic layer, growing of heteroepitaxial structure based on wide-gap III-nitrides on the basic layer and formation of the source, gate and drain. The heat conductive layer of CVD polycrystalline diamond is used as a pedestal; nickel is implanted to its surficial region and annealed. Before formation of the source, gate and drain an additional layer of insulating polycrystalline diamond and additional layers of hafnium dioxide and aluminium oxide are deposited on top of the transistor crystal; the total thickness of the above layers is 1.0-4.0 nm.

EFFECT: invention allows increased heat removal from the active part of SHF-transistor and minimisation of gate current losses.

6 cl, 4 dwg

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